JPS62138427A - Radiosensitizer - Google Patents

Abstract

PURPOSE:A radiosensitizer, containing a specific 3-nitro-1,2,4-triazole compound as an active ingredient, capable of remarkably increasing the sensitivity of hypoxic cells to radiation and increasing the radiotherapeutic effect with low toxicity. CONSTITUTION:A radiosensitizer containing a 3-nitro-1,2,4-triazole compound expressed by the formula {X is halogen, group expressed by the formula -CO-R1 [R1 is a group of -OR2 or -N(R3)R4]; R2 and R3 are H, alkyl, etc.; R4 is a group of R3, etc.; R is H, alkyl, hydroxyalkyl, etc.}. Examples of a specific compound include 2-(3'-nitro-5'-bromo-1',2',4'-triazol-1'-yl)acetic acid ethanolamide, etc. The compound expressed by the formula is capable of promoting inactivation of intractable hypoxic cells present in malignant tumor by irradiation.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a radiosensitizer that contains a specific nitrotriazole compound as an active ingredient, and which suppresses the damage caused by radiation irradiation to refractory hypoxic cells present in malignant tumors. This invention relates to a radiosensitizer that promotes activation.

Traditionally, radiation therapy, surgical therapy, chemotherapy, immunotherapy, etc. have been used to treat malignant tumors, and radiotherapy is an effective treatment method that has been used for many years. be.

However, there are problems in that it may not be cured even with radiotherapy, and that nrr+m may recur even if the eruption is cured.

The reasons for this include the radioresistance of the l1ffl tumor tissue itself and the presence of oxygen-deficient radioresistant cells in the tumor. In fact, in radiation irradiation experiments, it is known that cells in an oxygen-free atmosphere are two to three times more resistant to radiation than cells in the presence of oxygen.

Under these circumstances, there has been a strong demand for the development of hypoxic cell sensitizers, which are drugs that increase the sensitivity of hypoxic cells to radiation, as extremely effective means for improving the effects of radiation therapy.

From this point of view, several hypoxic cell aggravating agents have been developed, and for example, nido-10 imidazole derivatives are known as representative examples.

However, although misonibuzole, one of the representative compounds of nitroimidazole derivatives, exhibits a sensitizing effect approximately twice as high as that without additives in animal transplantation tumor experiments, it is difficult to administer large amounts due to neurotoxicity, and clinical No sensitizing effect has been recognized from the results obtained when the drug was administered to the human body using a suitable dosage method.

The present inventors conducted extensive research to find a compound with low toxicity and a higher sensitizing effect, and as a result, the following general formula (
I) The 3-21-rho 1,2,1I-1 riadur compound significantly increases the radiation response of hypoxic cells to radiation and increases the effectiveness of radiotherapy i') I found it.

(wherein, X represents a halogen atom or -Co-R,
R1 represents -0-R2 or -N(R,)R, R2
represents a hydrogen atom, an alkyl group, a hydroxyalkyl group, an alkyl group having an ether bond, or an alkyl group having a hydroxyl group and an ether bond, and R represents a hydrogen atom, an alkyl group, a hydroxyalgyl group, an alkyl group having an ether bond. or an alkyl group having a hydroxyl group and an ether bond, R4 is a group represented by R3, or -Rs-N (R6) R7, R2 is an alkylene group, and R6 and R1 are represented by R3 Furthermore, R1 and R4 or R6 and R7 may be bonded to each other to indicate an alkylene group or oxadialkylene group, and R3 and R6 may be bonded to each other to indicate an alkylene group.

R is a hydrogen atom, an alkyl group, a hydroxyalkyhalogen atom, an acyloxy group, -0-1h or -N(R3)-
Indicates R4. ) Hereinafter, the nitrotriazole compound having a specific substituent of the present invention will be described in detail.

In the above compounds, the alkyl group includes methyl, ethyl, propyl, isopropyl, butyl, isobutyl,
Nibutyl, amyl, hexyl, heptyl, octyl,
Examples include isooctyl, 2-ethylhexyl, etc., and hydroxyalkyl groups include 2-hydroxyethyl, 2-ethylhexyl, etc.
Examples of the alkyl group having an ether bond include methoxyethyl, ethoxyethyl, butoxyethyl, ethoxyethoxyethyl, etc., and examples of the alkyl group having a hydroxyl and ether bond include 2-(2” -hydroxyethoxy)ethyl, etc.

Examples of the alkylene group include methylene, ethylene, trimethylene, 1,2-propylene, tetramethylene, pentamethylene, 5-hexylene, 2,6-hebutylene, hexamethylene, and the like.

Examples of the oxadialkylene group include oxadi-diethylene and the like.

Examples of the acyl group include acetyl, propionyl, butyroyl, acryloyl, methacryloyl, hendyl, toluoyl, and the like.

Therefore, the compounds represented by the above general formula of the present invention include 2-(3'-2!-rho5"-bromo-1", 2"3
4″-triazol-1°-yl)acetic acid ethanolami)”, 2-(3°-21・rho5′-bromo-1′, 2
″, 4′-triazol-1″-yl)acetic acid jetanolamide, 2-G(′-2I-rho5′-bromo1′)
, 2', 4'-1-riazu-1°-yl) vinegar convex modified methoxyethylamide hydroxypropyl)-3-21-rho 5-bromo-1
, 2,4-1-riazole, ■-methyl-3-nitro 1.2.4-triazole-5-methylcarphonate,
3-nido L: 1-12゜4-1-riazole-5-carboxylic acid me[-xyethylamide, ■-methoxycarbonylmethyl-3-nitro-1゜2.4-triazole-
Methyl 5-carboxylate, L (3'-methoxy-2'-hydroxypropyl)-3-dito-1,2'4-
Methyl triazole-5-carboxylate, ■-methyl-3
-nitro 1.2.4-triazole-5-carboxylic acid ethanolamide, ■-(2°, 3'-epoxypropyl)-3-nitro 1.2.4-triazole-5-
Carboxylic acid ethanolamide, 1-(3”-methoxy-
2”-hydroxypropyl)-3-nitro-1,2,4
-) Riazole-5-carboxylic acid ethanolamide, 1
-(3°-acetoxy-2'-hydroxypropyl)-
3-nidoshi)-12,4-triazole-5-carboxylic acid ethanolamide, 1-(2'-hydroxyethyl)
-3-nitro-1,2,4-1-riazole-5-carboxylic acid ethanolamide, 1-(2°-hydroxyethylcarbamoylmethyl)-3-nitro-1°2.4-
Triazole-5-carboxylic acid ethanolamide, 3-
Nitro 1.2.1-triazole-5-carboxylic acid methoxyethylamide, l-methoxycarbonylmethyl-
3-Nitro-L2. f-Triff/-5-carboxylic acid metquinethylamide, 1-(2°-hydroxyethylcarbamoylmethyl)-3-21-17-1.2.4
-Triazole-5-carboxylic acid methoxyethylamide, 1- (N,N-bis(2”-hydroxyethyl)
Carbamoylmethyl]-3-nitro-1,2,4-1-
Riazole-5-carboxylic acid methoxyethylamide, ■
-(3°-aziridino-2"-hydroxypropyl)-
3-nitro-1,2,1-triazole-5-carboxylic acid methoxyethylamine i', 1-(2'-hydroxyethylcarbamoylethyl)-3-nitro-1,2,4-
triazole-J-carboxylic acid methoxyethylamide,
1-(Hydroxyethoxycarbonylmethyl)-3-nitro-1,2,4-1-riazole-5-carboxylic acid methoxyethylamide, 1-(ethoxyethoxycarbonylmethyl)-3-nitro-1,2゜4 -Triazole-5-carboxylic acid methoxyethylamide, 1-(hydroxyethoxyethoxycarbonylmethyl)-3-nitro-1,2,4-1-riazole-5-carboxylic acid methoxyethylamide, 1-methyl-3-di 1-Low L2.
=1-1-Riazole-5-carboxylic acid jetanolamide, ■-Methyl-3-2I-L2.4-triazole-5-carboxylic acid (2', 3'-dihydroxyph).

ropylamyl”, 1-(morpholinocarbonylmethyl)
-3-nitro-1.2.4-1-riazole-5-carboxylic acid morpholide, 1-(aziridinocarbonylmethyl)-3-nitro-1,2,4-1-riazole-5-
1-(Piperidinocarbonylmethyl)-3-2-2-carboxylic acid I”, 1-(piperidinocarbonylmethyl)-3-2-1.2.4-1-Riazole-5-carboxylic acid piperidide, 1-(4°-methylpiperazinocarbonylmethyl )-3-nitro-1,2,4-
1-Riazole-5-carboxylic acid (4”-methylbiveracito), 1-(diethylcarbamoylmethyl)-3-
Nitro-1,2,4-1-riazole-5-carboxylic acid diethylamide, i(dimethylaminopropylcarbamoylmethyl)-3-nitro-1,2°4-triazole-5-carboxylic acid dimethylaminopropylamide, ■-
Examples include (morpholinopropylcarbamoylmethyl)-3-nitro-1,2,4-1-riazole-5=carboxylic acid molporinopropylamide.

Moreover, when these compounds have an amino group, they may naturally be acid addition salts, and examples of acids that form this acid addition salt include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. inorganic acids and acetic acid, propionic acid, oxycarboxylic acid,
Examples include organic acids such as citric acid, hexatalic acid, adipic acid, lactic acid, and p-toluenesulfonic acid.

These compounds include, for example, 3-nitro-1,2,4-
A method of reacting a triazo-/re5-carboxylic acid ester or amide with a halocarboxylic acid ester or an unsaturated carboxylic acid ester, and then converting it into an amide if necessary; a method of reacting an epihalohydrin, and then reacting with a carboxylic acid, if necessary. Method of adding amines or alcohols: It can be produced by adding an alkylene oxide and then acylating if necessary.

Next, specific production examples of the compounds of the present invention will be described, but the present invention is not limited to these production examples.

Production Example 1 1.1 g of methyl 3-nitro-1,2,4-1-riazole-5-carboxylate was dimethylformaminated, 280 mg of sodium hydride was added, and the mixture was heated at room temperature for 20 min.
Stir for a minute. Thereafter, 1.3 g of methyl bromoacetate was added, and the mixture was stirred at 60 to 70°C for 4 hours. The solvent was distilled off under reduced pressure, and the mixture was extracted with ethyl acetate. Ethyl acetate was distilled off to obtain 1.1 g of a pale yellow oily product.

As a result of infrared spectroscopy, there was an ester-based absorption at 1740 cm-', confirming that it was the target product.

Elemental analysis 0%! (% N% Calculated value 34.4 3.28 23.0 Actual value
34.2 3.26 23.3 Production Example 2 530 mg of the compound obtained in Production Example 1 was dissolved in 2 ml of dioxane, and 500 + ng of monoethanolamine was dissolved.
was added and stirred at 70 to 80°C for 2 hours. After removing the solvent, it was treated with a cation exchange resin (manufactured by Dow: Dokai ax 50w), and then recrystallized from methanol/benzene to give a melting point of 10.
Colorless crystals at 6-108°C were obtained.

IR (KBr method): 3350, 3300, 1670, 1
560, 153o, 1320 and 1040 cm-' Elemental analysis 0% % N% Calculated value 36.0, i. 00 28.0 Actual value
35.8 3.91 27.9 Production Example 3 1 g of methyl 3-nitro-1.2.4-1-riazole-5-carboxylate was dissolved in 5 ml of dioxane, and methyl xyethylamine Ig was added. , 2 at 40-50”C
Stir for hours.

Excess amine and solvent were distilled off to obtain a colorless oily product.

IR: 3400, ■650, 1540 and 112
0 cm-' Elemental analysis 0% H%
N% calculated value 33.5 4.19 32.6
Actual value 33.8 4.20 32.3 Production Example 4 1.25 g of the compound obtained in Production Example 3 was dissolved in 20 ml of dimethylformamide, 280 μl of sodium hydride was added, and the mixture was stirred for 15 minutes. Methyl bromoacetate1.
3 g was added and stirred at 60-70°C for 2 hours. After the solvent was distilled off, the mixture was extracted with chloroform, and the chloroform was distilled off to obtain 1 g of a pale yellow oily product.

IR: 3350, 1750, 1690, 1560
, 1320, 1230 and 1130 cm-' Elemental analysis 0% f (% N% calculated value 37.6 4.53 24.4 actual value
37.8 4.55 24.7 Production Example 5 1-(N,N-bis(hydroxyethyl)carbamoylmethyl-3-nitro-1.2.4-1-riazole-5-produced in Production Example 4) Compound 530 1yo was dissolved in 3 ml of dioxane, 1 g of jetanolamine was added, and the mixture was stirred at 70 to 80°C for 2 hours. After distilling off the dioxane, a cation exchange resin (manufactured by Dow: Dowex 50) was added.
w) and chromatography on silica gel using chloroform/methanol as a developing solvent to obtain 380 cm of a colorless oily product.

IR: 3300-3450, 1650, 1550
, 131O, 1120 and 1070 cm-' Elemental analysis 0% 11% N% calculated value 40.0 5.56 23.3 Actual value
40.4 5.48 23.1 Production Example 6 ■-Methyl-3-nitro 1.2.4-) 470 ■ of methyl lyazole-5-carboxylate was dissolved in 3 ml of dioxane, and 3001 ■ of jetanolamine was added. .

After stirring at 60-70°C for 2 hours, dioxane was distilled off.
After treatment with an ion exchange resin (Dowex 50w), it was recrystallized from methanol/henganese to a melting point of I.
The product was obtained as a colorless solid with a temperature of II to 113°C.

IR (KBr method): 3300.1660.1560.1
31O and 1040 cm-' Elemental analysis 0% I (% N% calculated value 37.1 5.02 27.0 Actual 4) value
31.4 5.10 26.8 Production Example 7 The same procedure as Production Example 6 was carried out except that monoethanolamine was used instead of jetanolamine, with a melting point of 108 to 111.
“C colorless solid product was obtained.

IR (KBr method): 3450.3250.1690.1
560.1310 and 1080 cm- Elemental analysis 0% 8% N% calculated value
33.5 4.19 32.6 Actual value 33.
1 4.12 32.9 Production Example 8 Instead of diethanolamine, 2,3-dihydrokizib [
A colorless solid product with a melting point of 125-126°C was obtained in the same manner as in Production Example 6, except that cobylamine was used.

[R (KBr method): 3350.1690.1560
．． 1320 and 1040 cm- Elemental analysis 0% 14% N% calculated value 34.3 4.49 28.6 Actual value 33
．． 8 4.37 28.2 Production Example 9 Production 2 g of methyl 3-nitro-1,2,4-triazo'-5-carboxylate, 6 ml of methylglycidol and 0.2 g of anhydrous potassium carbonate were taken and heated at 80°C for 20 Stir for a minute.

After filtration, it was concentrated under residual pressure, and the residue was purified by silica gel chromatography using chloroform/methanol as a developing solvent to obtain 1.2 g of a colorless oily product.

IR: 3400.1740.1560.131O1
1240 and 1050 cm-' Elemental analysis 0% % N% Calculated value 36.9 4.62 21.5 Actual value 36
．． 4 4.75 21.7 Production Example 10 Dissolve compound 3501■ of Production Example 9 in dioxane 5+nl, process monoethanolamine 500 uvr, 8
Stirred at 0-100°C for 3 hours. Dioxane was distilled off, and a cation exchange resin (manufactured by Dow: Dowex 50w
), purified by silica gel chromatography using chloroform/methanol as a developing solvent, and purified with 0.3FS
A colorless oil product was obtained.

IR: 3300-3400.1675.1555.
1.1tlQ, 1120 and 1060 cm-' Elemental analysis profit 8% N day calculated value
37.4 5.19 24.2 Actual value 37.
1 5.12 23.9 Production Example 11 Same as Production Example 10 except that jetanolamine was used instead of monoethanolamine. A 11-color oily product was obtained.

IR: 3250-3400.1670.1555.
1320.1120 and 1055 cm-' Elemental analysis 0% 11% N% calculation a 39.6 5.71 21.
0'Actual value 39.8 5
．． 58 20.7 Production Example 12 L (2'-hydroxyethyl)-3-21-rho1.
2. , Preparation of 1-triazole-5-carboxylic acid ethanolamide 3-nitro1.2.4-1-riazole-
Methyl 5-carboxylate 3.0 g, ethylene oxide 1
．． 53 g of sodium methoxide was dissolved and stirred at 50-55°C for 1.5 hours. After distilling off the solvent and washing with Hensen, 1.94 g of colorless solid product was obtained.
I got it.

This product and 0.62 g of monoethanolamine were dissolved in 10 ml of dioxane and stirred at 80-100°C for 3 hours. Soo; 1-san was distilled off, treated with a cation exchange resin (Dowex 50w), and purified by silica gel chromatography using chloroform/methanol as a developing solvent to obtain a colorless solid with a melting point of 108 to 110°C. The product was obtained.

IR: 3400, 3250, 1670, 1555
, 1320 and 1060 cm-' Elemental analysis 0% 8% N% calculated value
34.3 4.49 28.6 Actual value 3
4.0 4.55 28.8 The above compounds of the present invention are useful as sensitizers in radiotherapy, and the dosage varies depending on the type of tumor and the compound, but in general, oral administration is 20 to 10,000 mg. , 0.5 to 10,000 mg for injections, 20 to 10 for suppositories
000 mg, and the optimal dose is determined based on the doctor's judgment according to the symptoms, depending on the type of radiation, the amount of irradiation, the degree of fractionation of irradiation, etc.

Furthermore, there are no particular restrictions on the dosage form of the compound of the present invention (,
As carriers, those commonly used in the pharmaceutical field can be used.
Eight rounds are made according to methods commonly used in this field.

The effects of radiation aggravation of the compounds of the present invention will be shown below using specific examples.

Example = 1 ① In order to examine the radiosensitizing effect on -79 Chinese hamster cells, 10,000 C.79 cells were cultured in a monolayer in a glass petri dish, and V-79 cells for logarithm were prepared.

A medium solution of a test compound at a predetermined concentration was added to a Petri dish, left to stand at 37°C for 60 minutes, then placed in a sealed container at room temperature, nitrogen gas was passed through it for 10 minutes to eliminate oxygen, and L. 6
X-rays were irradiated at a radiation rate of Gy/min.

After irradiation, wash with phosphate buffer, make single cells with trypsin, put the specified amount into a culture dish, and add medium 5+nl.
was added and cultured at 37°C for 7 days, washed with water after staining, and the number of colonies formed was measured.

For comparison, tests were also conducted in which only a medium solution containing no compound was added and irradiated under nitrogen and irradiated in the presence of air at a=t.

By calculating the cell survival rate from these values and plotting the logarithm of the survival rate against the irradiation dose, a linear relationship is obtained.

Find the intersection of this straight line and the horizontal straight line where the survival rate is 1.0, and calculate the interlead open dose: Dq (Gy), and the irradiation dose required to reduce the survival rate to 1710 from the slope of the straight line:
D+o (Gy) was determined.

In addition, the radiation M (Do. , , , = l) q + 3 0 + o) required to inactivate 99.9% of the cells was determined, and the ratio to the value of irradiation under air irradiation (Do. + y ./I
) o. +. /． ), and calculate the nitrogen reference sensitization ratio (
SARA number) and nitrogen reference sensitization ratio (N2 group (1ξSARA number)
RA number) was set as a constant.

The results obtained are shown in Table 1.

Example-2 105 EMT-6 tumor cells were inoculated into the thigh skin F of both legs of Balb/C male mice (8 weeks old, 4 mice per group). After tumor cell inoculation, when the tumor size reaches approximately 1 cm in diameter, a physiological saline solution of the test compound is administered intraperitoneally.
mg/kg), 450rad after 40 minutes, X in 7 minutes
irradiation, and mice were killed 5 minutes after irradiation.

After whole body sterilization with 70% ethanol, the tumor area was excised, the tissue was cut into small pieces, mixed with 22 ml of trypsin, and stirred in T:PA at 37°C for 50 minutes. Remove the supernatant, count the number of cells, and place the designated chamber in a plastic plate 1 with a diameter of 5 cm.
After adding 5 ml of medium, the cells were cultured in a carbon dioxide incubator. Cells that had not been irradiated with X-rays were removed from the incubator after 9 days, and cells that had been irradiated with X-rays were removed from the incubator after 10 days. Fix the cells, stain the cells with Giemsa staining solution, and count the number of colonies formed.

Cells that were not irradiated with X-rays were used as a control, and the survival rate was measured. The results are shown in Table-2.

Claims (1)

[Claims] 3-nitro-1,2,4 represented by the following general formula (I)
- containing a triazole compound as an active ingredient,
Radiosensitizer. ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (I) (In the formula, X represents a halogen atom or -CO-R_1, R_1 represents -O-R_2 or -N(R_3)R_4, R_2 represents a hydrogen atom, An alkyl group, a hydroxyalkyl group, an alkyl group having an ether bond, or an alkyl group having a hydroxyl group and an ether bond, R
_3 is a hydrogen atom, an alkyl group, a hydroxyalkyl group,
Represents an alkyl group having an ether bond or a hydroxyl group and an alkyl group having an ether bond, R_4 is a group represented by R_3, or -R_5-N(R_6)R
_7, R_5 represents an alkylene group, R_6 and R_7 represent the group represented by R_3, and R_3 and R_
4 or R_6 and R_7 may be combined with each other to represent an alkylene group or an oxadialkylene group, and further R_
3 and R_6 may be bonded to each other to represent an alkylene group. R is a hydrogen atom, an alkyl group, a hydroxyalkyl group,
▲There are mathematical formulas, chemical formulas, tables, etc.▼, -CH_2-CH
(OH)-CH_2-Y or R_8-CO-R_1, R_8 represents an alkylene group, Y is a halogen atom, an acyloxy group, -O-R_2 or -N(R_3)-R
Shows _4. )